Fractional 13C enrichment of isolated carbons using [1-13C]- or [2- 13C]-glucose facilitates the accurate measurement of dynamics at backbone Calpha and side-chain methyl positions in proteins - PubMed (original) (raw)

. 2007 Jul;38(3):199-212.

doi: 10.1007/s10858-007-9158-6. Epub 2007 Jun 7.

Affiliations

Fractional 13C enrichment of isolated carbons using [1-13C]- or [2- 13C]-glucose facilitates the accurate measurement of dynamics at backbone Calpha and side-chain methyl positions in proteins

Patrik Lundström et al. J Biomol NMR. 2007 Jul.

Abstract

A simple labeling approach is presented based on protein expression in [1-(13)C]- or [2-(13)C]-glucose containing media that produces molecules enriched at methyl carbon positions or backbone C(alpha) sites, respectively. All of the methyl groups, with the exception of Thr and Ile(delta1) are produced with isolated (13)C spins (i.e., no (13)C-(13)C one bond couplings), facilitating studies of dynamics through the use of spin-spin relaxation experiments without artifacts introduced by evolution due to large homonuclear scalar couplings. Carbon-alpha sites are labeled without concomitant labeling at C(beta) positions for 17 of the common 20 amino acids and there are no cases for which (13)C(alpha)-(13)CO spin pairs are observed. A large number of probes are thus available for the study of protein dynamics with the results obtained complimenting those from more traditional backbone (15)N studies. The utility of the labeling is established by recording (13)C R (1rho) and CPMG-based experiments on a number of different protein systems.

PubMed Disclaimer

References

    1. Methods Enzymol. 1994;239:563-96 - PubMed
    1. J Biomol NMR. 1996 Mar;7(2):157-62 - PubMed
    1. J Am Chem Soc. 2003 Nov 26;125(47):14324-35 - PubMed
    1. J Am Chem Soc. 2001 Nov 21;123(46):11341-52 - PubMed
    1. J Am Chem Soc. 2002 Feb 20;124(7):1443-51 - PubMed

Publication types

MeSH terms

Substances

LinkOut - more resources